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Journal of Histochemistry and Cytochemistry, Vol. 50, 11-20, January 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

Ultrastructural Staining with Sodium Metaperiodate and Sodium Borohydride

Maria V.T. Loboa, F. Javier M. Alonsoa, Maria I. Arenasa, Enrique Casoa, Benito Frailea, and Rafael Martín del Ríoa
a Servicio de Neurobiología, Departamento de Investigación, Laboratorio de Oncologia Molecular Applicada, Departamento de Oncologia Médica, Hospital Ramón y Cajal, and Departamento de Biología Celular y Genética, Universidad de Alcala, Alcala de Henares, Madrid, Spain

Correspondence to: Maria V.T. Lobo, Servicio de Neurobiología, Departamento de Investigación, Hospital Ramón y Cajal, Ctra. de Colmenar Km 9, 28034 Madrid, Spain. E-mail: rafael.martin@hrc.es

This article describes new ultrastructural staining methods for osmicated tissues based on the incubation of sections with sodium metaperiodate and sodium borohydride solutions before uranyl/lead staining. Sections incubated with sodium metaperiodate and sodium borohydride, treated with Triton X-100, and stained with ethanolic uranyl acetate/lead citrate showed a good contrast for the nucleolus and the interchromatin region, whereas the chromatin masses were bleached. Chromatin bleaching depended on the incubation with these oxidizing (metaperiodate) and reducing (borohydride) agents. Other factors that influenced the staining of the chromatin masses were the en bloc staining with uranyl acetate, the incubation of sections with Triton X-100, and the staining with aqueous or ethanolic uranyl acetate. The combination of these factors on sections treated with metaperiodate/borohydride provided a different appearance to the chromatin, from bleached to highly contrasted. Most cytoplasmic organelles showed a similar appearance with these procedures than with conventional uranyl/lead staining. However, when sections were incubated with metaperiodate/borohydride and Triton X-100 before uranyl/lead staining, the collagen fibers, and the glycocalix and zymogen granules of pancreatic acinar cells, appeared bleached. The possible combination of these methods with the immunolocalization of the amino acid taurine was also analyzed. (J Histochem Cytochem 50:11–19, 2002)

Key Words: ultrastructural staining, cytochemical methods, immunocytochemistry


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